LOCUS       sequence-181755-        6934 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module A, Promoter: CmYLCV, Gene: AtCas9, Terminator: HSP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6934)
  AUTHORS   Cermak T, Curtin SJ, Gil-Humanes J, Cegan R, Kono TJY, Konecna E, 
            Belanto JJ, Starker CG, Mathre JW, Greenstein RL, Voytas DF
  TITLE     A multi-purpose toolkit to enable advanced genome engineering in 
            plants.
  JOURNAL   Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 
            10.1105/tpc.16.00922.
  PUBMED    28522548
REFERENCE   2  (bases 1 to 6934)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant Cell.
            2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922."
COMMENT     Sequence Label: pMOD_A0103
FEATURES             Location/Qualifiers
     source          1..6934
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      80..668
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     569..588
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     822..839
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             1368..5471
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             1368..5471
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5484..5504
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5517..5765
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     promoter        5842..5946
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gccttgacgg caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
       61 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
      121 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ctaactcttt
      181 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
      241 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
      301 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
      361 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgctcacagc
      421 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
      481 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
      541 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
      601 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
      661 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
      721 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
      781 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
      841 aagcggaaga gcgcccacct gctcatgatc ggcgcgcctg gcagacatac tgtcccacaa
      901 atgaagatgg aatctgtaaa agaaaacgcg tgaaataatg cgtctgacaa aggttaggtc
      961 ggctgccttt aatcaatacc aaagtggtcc ctaccacgat ggaaaaactg tgcagtcggt
     1021 ttggcttttt ctgacgaaca aataagattc gtggccgaca ggtgggggtc caccatgtga
     1081 aggcatcttc agactccaat aatggagcaa tgacgtaagg gcttacgaaa taagtaaggg
     1141 tagtttggga aatgtccact cacccgtcag tctataaata cttagcccct ccctcattgt
     1201 taagggagca aaatctcaga gagatagtcc tagagagaga aagagagcaa gtagcctaga
     1261 agtagtcaag gcggcgaagt attcaggcac gtggccagga agaagaaaag ccaagacgac
     1321 gaaaacaggt aagagctaag cttcctgcag gtagatcgct cgtcgacatg gataagaagt
     1381 actctatcgg actcgatatc ggaactaact ctgtgggatg ggctgtgatc accgatgagt
     1441 acaaggtgcc atctaagaag ttcaaggttc tcggaaacac cgataggcac tctatcaaga
     1501 aaaaccttat cggtgctctc ctcttcgatt ctggtgaaac tgctgaggct accagactca
     1561 agagaaccgc tagaagaagg tacaccagaa gaaagaacag gatctgctac ctccaagaga
     1621 tcttctctaa cgagatggct aaagtggatg attcattctt ccacaggctc gaagagtcat
     1681 tcctcgtgga agaagataag aagcacgaga ggcaccctat cttcggaaac atcgttgatg
     1741 aggtggcata ccacgagaag taccctacta tctaccacct cagaaagaag ctcgttgatt
     1801 ctactgataa ggctgatctc aggctcatct acctcgctct cgctcacatg atcaagttca
     1861 gaggacactt cctcatcgag ggtgatctca accctgataa ctctgatgtg gataagttgt
     1921 tcatccagct cgtgcagacc tacaaccagc ttttcgaaga gaaccctatc aacgcttcag
     1981 gtgtggatgc taaggctatc ctctctgcta ggctctctaa gtcaagaagg cttgagaacc
     2041 tcattgctca gctccctggt gagaagaaga acggactttt cggaaacttg atcgctctct
     2101 ctctcggact cacccctaac ttcaagtcta acttcgatct cgctgaggat gcaaagctcc
     2161 agctctcaaa ggatacctac gatgatgatc tcgataacct cctcgctcag atcggagatc
     2221 agtacgctga tttgttcctc gctgctaaga acctctctga tgctatcctc ctcagtgata
     2281 tcctcagagt gaacaccgag atcaccaagg ctccactctc agcttctatg atcaagagat
     2341 acgatgagca ccaccaggat ctcacacttc tcaaggctct tgttagacag cagctcccag
     2401 agaagtacaa agagattttc ttcgatcagt ctaagaacgg atacgctggt tacatcgatg
     2461 gtggtgcatc tcaagaagag ttctacaagt tcatcaagcc tatcctcgag aagatggatg
     2521 gaaccgagga actcctcgtg aagctcaata gagaggatct tctcagaaag cagaggacct
     2581 tcgataacgg atctatccct catcagatcc acctcggaga gttgcacgct atccttagaa
     2641 ggcaagagga tttctaccca ttcctcaagg ataacaggga aaagattgag aagattctca
     2701 ccttcagaat cccttactac gtgggacctc tcgctagagg aaactcaaga ttcgcttgga
     2761 tgaccagaaa gtctgaggaa accatcaccc cttggaactt cgaagaggtg gtggataagg
     2821 gtgctagtgc tcagtctttc atcgagagga tgaccaactt cgataagaac cttccaaacg
     2881 agaaggtgct ccctaagcac tctttgctct acgagtactt caccgtgtac aacgagttga
     2941 ccaaggttaa gtacgtgacc gagggaatga ggaagcctgc ttttttgtca ggtgagcaaa
     3001 agaaggctat cgttgatctc ttgttcaaga ccaacagaaa ggtgaccgtg aagcagctca
     3061 aagaggatta cttcaagaaa atcgagtgct tcgattcagt tgagatttct ggtgttgagg
     3121 ataggttcaa cgcatctctc ggaacctacc acgatctcct caagatcatt aaggataagg
     3181 atttcttgga taacgaggaa aacgaggata tcttggagga tatcgttctt accctcaccc
     3241 tctttgaaga tagagagatg attgaagaaa ggctcaagac ctacgctcat ctcttcgatg
     3301 ataaggtgat gaagcagttg aagagaagaa gatacactgg ttggggaagg ctctcaagaa
     3361 agctcattaa cggaatcagg gataagcagt ctggaaagac aatccttgat ttcctcaagt
     3421 ctgatggatt cgctaacaga aacttcatgc agctcatcca cgatgattct ctcaccttta
     3481 aagaggatat ccagaaggct caggtttcag gacagggtga tagtctccat gagcatatcg
     3541 ctaacctcgc tggatctcct gcaatcaaga agggaatcct ccagactgtg aaggttgtgg
     3601 atgagttggt gaaggtgatg ggaaggcata agcctgagaa catcgtgatc gaaatggcta
     3661 gagagaacca gaccactcag aagggacaga agaactctag ggaaaggatg aagaggatcg
     3721 aggaaggtat caaagagctt ggatctcaga tcctcaaaga gcaccctgtt gagaacactc
     3781 agctccagaa tgagaagctc tacctctact acctccagaa cggaagggat atgtatgtgg
     3841 atcaagagtt ggatatcaac aggctctctg attacgatgt tgatcatatc gtgccacagt
     3901 cattcttgaa ggatgattct atcgataaca aggtgctcac caggtctgat aagaacaggg
     3961 gtaagagtga taacgtgcca agtgaagagg ttgtgaagaa aatgaagaac tattggaggc
     4021 agctcctcaa cgctaagctc atcactcaga gaaagttcga taacttgact aaggctgaga
     4081 ggggaggact ctctgaattg gataaggcag gattcatcaa gaggcagctt gtggaaacca
     4141 ggcagatcac taagcacgtt gcacagatcc tcgattctag gatgaacacc aagtacgatg
     4201 agaacgataa gttgatcagg gaagtgaagg ttatcaccct caagtcaaag ctcgtgtctg
     4261 atttcagaaa ggatttccaa ttctacaagg tgagggaaat caacaactac caccacgctc
     4321 acgatgctta ccttaacgct gttgttggaa ccgctctcat caagaagtat cctaagctcg
     4381 agtcagagtt cgtgtacggt gattacaagg tgtacgatgt gaggaagatg atcgctaagt
     4441 ctgagcaaga gatcggaaag gctaccgcta agtatttctt ctactctaac atcatgaatt
     4501 tcttcaagac cgagattacc ctcgctaacg gtgagatcag aaagaggcca ctcatcgaga
     4561 caaacggtga aacaggtgag atcgtgtggg ataagggaag ggatttcgct accgttagaa
     4621 aggtgctctc tatgccacag gtgaacatcg ttaagaaaac cgaggtgcag accggtggat
     4681 tctctaaaga gtctatcctc cctaagagga actctgataa gctcattgct aggaagaagg
     4741 attgggaccc taagaaatac ggtggtttcg attctcctac cgtggcttac tctgttctcg
     4801 ttgtggctaa ggttgagaag ggaaagagta agaagctcaa gtctgttaag gaacttctcg
     4861 gaatcactat catggaaagg tcatctttcg agaagaaccc aatcgatttc ctcgaggcta
     4921 agggatacaa agaggttaag aaggatctca tcatcaagct cccaaagtac tcactcttcg
     4981 aactcgagaa cggtagaaag aggatgctcg cttctgctgg tgagcttcaa aagggaaacg
     5041 agcttgctct cccatctaag tacgttaact ttctttacct cgcttctcac tacgagaagt
     5101 tgaagggatc tccagaagat aacgagcaga agcaactttt cgttgagcag cacaagcact
     5161 acttggatga gatcatcgag cagatctctg agttctctaa aagggtgatc ctcgctgatg
     5221 caaacctcga taaggtgttg tctgcttaca acaagcacag agataagcct atcagggaac
     5281 aggcagagaa catcatccat ctcttcaccc ttaccaacct cggtgctcct gctgctttca
     5341 agtacttcga tacaaccatc gataggaaga gatacacctc taccaaagaa gtgctcgatg
     5401 ctaccctcat ccatcagtct atcactggac tctacgagac taggatcgat ctctcacagc
     5461 tcggtggtga ttcaagggct gatcctaaga agaagaggaa ggtttgacgt cgacgatatg
     5521 aagatgaaga tgaaatattt ggtgtgtcaa ataaaaagct tgtgtgctta agtttgtgtt
     5581 tttttcttgg cttgttgtgt tatgaatttg tggctttttc taatattaaa tgaatgtaag
     5641 atcacattat aatgaataaa caaatgtttc tataatccat tgtgaatgtt ttgttggatc
     5701 tcttctgcag catataacta ctgtatgtgc tatggtatgg actatggaat atgattaaag
     5761 ataagccaga gctctggtga cggacgtggg caggtgtgat gaaaagcctt agggtcaggt
     5821 ggcacttttc ggggaaatgt gcgatgaacc cctatttgtt tatttttcta aatacattca
     5881 aatatgtatc cgctaatgag acaataaccc tgataaatgc ttcaattata ttgaaaaagg
     5941 aagagtatga gtatccaaca tttccgtgtc gcccttattc ccttttttgc ggccttctgc
     6001 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     6061 ggtgcccgag tgggttacat tgaactggat ctcaacagcg gcaaaatctt agagagtttt
     6121 cgccccgaag aacgcttccc aatgatgagc actttcaaag ttctgctatg tggcgcggtg
     6181 ttatcccgta ttgatgctgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     6241 gacttggttg agtattcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     6301 gaattatgta gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     6361 acgattggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     6421 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     6481 acgatgcctg tagcaatggc aacaactctc cgcaaactat taactggcga actacttact
     6541 cttgcttcac gccagcaact cattgactgg atggaggcgg acaaagttgc aggaccactt
     6601 ctgcgctcgg cacttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     6661 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     6721 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6781 ggtgcctcac tgattaagca ttggtaactg tcagacgaag ggaaataaat agtagcccgc
     6841 ctgatgtgcg ggcttttttt tggtggtggt taccagtgtc gcttagctac cgtaggtcct
     6901 cttaggcggt ccgattagcg acatagtcat caat
//
